title: Paracrine factors of vascular endothelial cells facilitate cardiomyocyte differentiation of mouse embryonic stem cells.
abstract: For myocardial regeneration therapy, the low differentiation capability of functional cardiomyocytes sufficient to replace the damaged myocardial tissue is one of the major difficulties. Using Nkx2.5-GFP knock-in ES cells, we show a new efficient method to obtain cardiomyocytes from embryonic stem (ES) cells. The proportion of GFP-positive cells was significantly increased when ES cells were cultured with a conditioned medium from aortic endothelial cells (ECs), accompanied by upregulation of cardiac-specific genes as well as other mesodermal genes. The promotion was more prominent when EC-conditioned medium was added at an early stage of ES cell differentiation culture (Day 0-3). Inhibitors of bone morphogenic protein (BMP), cyclooxygenase (COX), and nitric oxide synthetase (NO) prevented the promotion of cardiomyogenesis by EC-conditioned medium. These results suggest that supplementation of EC-conditioned medium enables cardiomyocytes to be obtained efficiently through promotion of mesoderm induction, which is regulated by BMP, COX, and NOS.
authors.affiliation: Department of Cardiothoracic Surgery, Nagoya University, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550, Japan.
authors.forename: Miwako
authors.initials: M
authors.lastname: Kado
authors.affiliation: Department of Cardiovascular Research, Research Institute of Environmental Medicine, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi 464-8601, Japan. Electronic address: jlee@riem.nagoya-u.ac.jp.
authors.forename: Jong-Kook
authors.initials: JK
authors.lastname: Lee
authors.affiliation: Department of Bioscience, Research Institute, National Cardiovascular Center, 5-7-1 Fujishiro-dai, Suita, 565-8565, Japan.
authors.forename: Kyoko
authors.initials: K
authors.lastname: Hidaka
authors.affiliation: Department of Cardiovascular Research, Research Institute of Environmental Medicine, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi 464-8601, Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Miwa
authors.affiliation: Department of Cardiology, Graduate School of Medicine, Nagoya University, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550, Japan.
authors.forename: Toyoaki
authors.initials: T
authors.lastname: Murohara
authors.affiliation: Department of Pathology, Aichi Medical University, Nagakute-cho, Aichi, 480-1195, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Kasai
authors.affiliation: Department of Pathology, Aichi Medical University, Nagakute-cho, Aichi, 480-1195, Japan.
authors.forename: Shinsuke
authors.initials: S
authors.lastname: Saga
authors.affiliation: Department of Bioscience, Research Institute, National Cardiovascular Center, 5-7-1 Fujishiro-dai, Suita, 565-8565, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Morisaki
authors.affiliation: Department of Cardiothoracic Surgery, Nagoya University, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550, Japan.
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Ueda
authors.affiliation: Department of Cardiovascular Research, Research Institute of Environmental Medicine, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi 464-8601, Japan.
authors.forename: Itsuo
authors.initials: I
authors.lastname: Kodama
mesh_terms: D047228:Angiotensin II Type 1 Receptor Blockers / Q000494:pharmacology; D000818:Animals; D001011:Aorta / Q000166:cytology; D019485:Bone Morphogenetic Proteins / Q000037:antagonists & inhibitors; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D017077:Culture Media, Conditioned / Q000378:metabolism / Q000494:pharmacology; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D019332:Endothelin-1 / Q000037:antagonists & inhibitors; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D055879:Gene Knock-In Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins / Q000235:genetics; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000187:drug effects; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D011451:Prostaglandin-Endoperoxide Synthases / Q000187:drug effects; D044140:Receptor, Angiotensin, Type 1 / Q000187:drug effects; D014157:Transcription Factors / Q000235:genetics
chemical_list: D047228:Angiotensin II Type 1 Receptor Blockers; D019485:Bone Morphogenetic Proteins; D017077:Culture Media, Conditioned; D016861:Cyclooxygenase Inhibitors; D019332:Endothelin-1; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins; C484469:Nkx2-5 protein, mouse; D044140:Receptor, Angiotensin, Type 1; D014157:Transcription Factors; D049452:Green Fluorescent Proteins; D019001:Nitric Oxide Synthase; D011451:Prostaglandin-Endoperoxide Synthases